Through a career development plan which includes structured didactics, multidisciplinary mentoring and innovative research experience, the applicant will acquire skills and knowledge required as an independent investigator. The applicant will gain the expertise in proteomics and glial cell biology, as applicable to a career as a neuroscientist. Together with the proposed research experience, this will position her as a future leader in the field of heritable white matter disorders. The research plan aims to identify pathophysiological mechanisms and explore therapeutic targets in Vanishing White Matter (VWM) disease, a leukodystrophy caused by mutations in the eukaryotic translation initiation factor 2B (elF2B) gene. New preliminary data indicate that elF2B mutated cells display an abnormal accumulation of proteins within the endoplasmic reticulum (ER) as a response to stress. We hypothesize that abnormal protein trafficking, as a result of abnormally regulated ER stress, occurs in VWM, and is largely responsible for the phenotype of progressive myelin destruction in this disorder. The proposed aims are to: 1) define protein trafficking alterations in glial cells affected by elF2B mutations or RNA interference using a comparative proteomics approach (SILAC) combined with mass spectrometry to identify proteins with abnormal ER storage and secretion as a function of a number of chemical and physical stressors 2) determine the effect of chaperones on protein trafficking in elF2B mutated CMS cells by monitoring the effect of these candidate therapeutic molecules on the proteins identified in Aim 1. We will initially perform these studies on elF2B knockdown oligodendrocytes, and subsequently validate this paradigm on CNS cells obtained from a mouse model of VWM disease. The therapeutic effect of 4-phenylbutyrate (chemical chaperone), Salubrinal (translation factor inhibitor), and a select group of molecules identified by high throughput screening in elF2B mutated yeast, will be tested for ameliorating effect on stress response in VWM models. Agents (drugs) identified as effective in vitro will be tested in the mouse model and ultimately in patients affected by VWM as a long-term goal of this research. This research goal is consistent with the career goal of the applicant to improve understanding and treatment of patients with leukodystrophy.